I’m working on a science project and need support to help me understand better.
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Result for the project: Use this space to summarize the outcome of there search articles. Mention here what did the group learned through this project.
Discussion: In this space I want each participant to express in their own words the knowledge obtained from this project and how would they apply it as histotechnologists.
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The Demonstration of Melanin Pigment and Argentaffin Granules Using the Fontana Masson Staining Method Result
Abstrast: Melanin pigments and argentaffin granules are essential compounds in identifying melanocytic lesions, carcinoid tumors, and other medical conditions for a histopathologic diagnosis. Since melanin is barely detectable in unstained sections of the human epidermis, these substances tend to have a nonspecific visualization with routine hematoxylin-eosin (H&E) staining. Because of this, the Fontana-Masson (FM) staining method is used in histology for its specialized identification of melanin and argentaffin granules (Carriel, 2011). This technique demonstrates the ability of these granules to bind silver, consequently resulting in staining the melanin and argentaffin a brown-black color. In our presentation, we will observe the Fontana-Masson technique and the staining chemistry of how these argentaffin substances are identified. Additionally, we will exhibit the staining patterns that may be useful in cases of carcinoid tumors, melanoma, and fungal infections.
Introduction: Melanin pigments play a key role in determining human skin color and pigmentation. In the basal layer of the epidermis, melanin is synthesized by melanocytes and then transferred to surrounding keratinocytes. Argentaffin, a property of melanin, has the capacity to reduce metallic solutions which gives it the ability to take up silver staining. In the histology laboratory, routine H&E staining is nonspecific for identifying melanin pigments especially when the pigment is not abundant or other pigments are present in that tissue sample. Because of this, special stain techniques are utilized, such as ferrous iron, Schmorls method, WarthinStarry stain, and most notably the Fontana-Masson (FM) method. The FM technique is most often used in histopathology for melanin identification. This method works by reducing ammoniacal silver nitrate to metallic silver, producing an insoluble black-brown color under a light microscope. Identification and localization of melanin or other argentaffin granules can be an important component for diagnosing serious conditions such as melanocytic lesions (melanoma, etc.) or carcinoid tumors (Carriel, 2011). Due to the presence of melanin in the cell wall of certain fungi, the FM stain is applied to demonstrate particular fungal infections as well (Sundaram, 2014).
Material and method: Purpose: To demonstrate argentaffin granules of carcinoid tumors and melanin.
Reagents: Silver nitrate, 10% solution- 10 g silver nitrate and 100 mL distilled water.
-Fontana silver solution- 25 mL of silver solution mixed with ammonium hydroxide.
-Gold chloride, 0.2% solution- 10 mL gold chloride 1% solution with 40 mL distilled water.
-Sodium thiosulfate, 5% solution- 25 g sodium thiosulfate and 500 mL distilled water.
-Nuclear Fast Red
- Retrieve tissue sample fixed in 10% formalin, processed and embedded.
- Cut paraffin sections 4-5 ?m and place on slide.
- Remove paraffin and hydrate sections in distilled water.
- Immerse slides in silver nitrate solution for 1 hour in water bath (56C). Check slides for reaction which will be dark brown granules and a colorless background. Adjust time as needed for reaction.
- Rinse slides in distilled water.
- Immerse slides in Gold chloride, 0.2% solution for 10 minutes.
- Rinse slides in distilled water
REFERENCES : .Bishop J.A., Nelson A.M., Merz W.G., Askin F.B., Riedel S. Evaluation of the detection of melanin by theFontana-Masson silver stain in tissue with a wide range of organisms including Cryptococcus. Hum Pathol. 2012 Jun;43(6):898-903. doi: 10.1016/j.humpath.2011.07.021. Epub 2011 Dec 9.Carriel V.S., Aneiros-Fernandez J, Arias-Santiago S, Garzón IJ, Alaminos M, Campos A. A novelhistochemical method for a simultaneous staining of melanin and collagen fibers. J Histochem Cytochem. 2011 Mar;59(3):270-7. doi: 10.1369/0022155410398001. Epub 2011 Jan 12.Carson, F. L., Cappellano, C. (2020). In Histotechnology: A Self-Instructional Text (5th ed.). Chicago, Illinois: American Society for Clinical Pathology.Joly, T. N., Wibawa, J. I. D., Bell, M., & Tobin, D. (2016). Melanin fate in the human epidermis: a reassessment of how best to detect and analyse histologically. Experimental Dermatology, 25(7), 501504. https://doi.org/10.1111/exd.13016Sundaram C, Shantveer G U, Umabala P, Lakshmi V. Diagnostic utility of melanin production by fungi:Study on tissue sections and culture smears with Masson-Fontana stain. Indian J Pathol Microbiol 2014;57:217-22